CN110152013A - A kind of pectin-adriamycin conjugates and its preparation method and application - Google Patents
A kind of pectin-adriamycin conjugates and its preparation method and application Download PDFInfo
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Abstract
The present invention relates to a kind of pectin-adriamycin conjugates, and disclose the Preparation method and use of above-mentioned conjugates.Pectin of the invention-adriamycin conjugates has completely new chemical structure, can the accumulation for a long time of targeting high concentration in malignant tumor tissue, achieving the purpose that Synergy and attenuation, indication is the chemotherapy of various solid malignants.
Description
Technical field
The present invention relates to pharmaceutical fields, more particularly to a kind of pectin-adriamycin conjugates and its preparation method and application.
Background technique
" pectin-adriamycin conjugates " targeted drug delivery system (PAC) is in a large amount of previous works and literature research
On the basis of, select carrier of the pectin as high molecular anticarcinogenic prodrug, and modified by the structure to pectin, allow to and
Polypeptide-adriamycin covalent bond forms the water-soluble pectin with high drug load-adriamycin conjugates.
PAC is macromolecule conjugates, and partial size can benefit into some drugs of the circulatory system after 200nm or so, injection
The enhancing having with tumor tissues to macromolecular substances is penetrating and retention effect (EPR) and accumulates in tumor tissues, reach by
Moving-target to purpose.PAC is gradually swallowed by tumour cell in tumor tissues, and hydrolysis releases adriamycin in lysosome, is sent out
Wave the effect of killing tumour.Existing pectin-adriamycin conjugates be pectin and adriamycin directly by amido bond bonding or
The bonding of acylhydrazone key, is insoluble in water, it is necessary to it is made into nano suspension or freeze-dried powder, it can only local application；If systemic administration, easily
By reticuloendothelial system phagocytic, targeting is limited, can not high concentration accumulation for a long time in malignant tumor tissue, antitumor curative effect
It is bad.
Summary of the invention
It being capable of the accumulation for a long time of targeting high concentration in view of the above-mentioned deficiencies in the prior art, it is an object of the present invention to provide one kind
In the pectin in malignant tumor tissue-adriamycin conjugates, the purpose of Synergy and attenuation can reach, the present invention also provides above-mentioned fruits
Glue-adriamycin conjugates preparation method.
The present invention to achieve the above object the technical solution adopted is that: a kind of pectin-adriamycin conjugates, have such as formula
(I) structure:
a-b-c-d-e (I)
Wherein a is pectin or modifying pectin, and b PEGn, c are enzymatic lysis group, polypeptide or polypeptide derivative, and d is
Self-degradation group, e are adriamycin.
Further, the pectin is polygalacturonase without ester pectin, and the modifying pectin is poly galacturonic
The C that carbonyl of the acid without ester pectin and hydroxyl replace2-C4Alkylamine generate amide, be further activated as pectin carbonic ester；
Preferably, a is modifying pectin；Further preferably, the molecular weight of a is 1-4KD；Further preferably, the polygalacturonase
The galacturonic acid content of no ester pectin is 95% or more.
Further, in formula (I), n is the integer of 1-20, and c is dipeptides, dipeptidase derivant, tripeptides, tripeptide derivative, four
Peptide, tetrapeptide derivative；Preferably, n is the integer of 6-12；Preferably, c val-ala.
Further, d is to aminobenzyloxycarbonyl (PABC).
Further, the pectin is the C that carbonyl and hydroxyl replace2-C4The connected pectin of alkyl amine group；Preferably, institute
Stating pectin molecule amount is 1-4KD.
The invention also discloses pectin as described above-adriamycin conjugates preparation method, by formula (13) compound with
It is reacted under the conditions of formula (9) compound is existing for the alkaline reagent；
In formula (13), a is the integer selected from 2-4, m 1-60KD；Formula (9) compound is b-c-d-ADM, and ADM is Ah mould
Element, b PEGn, c are polypeptide；Preferably, c is dipeptides, dipeptidase derivant, tripeptides, tripeptide derivative, tetrapeptide, tetrapeptide derivative；
Preferably, n is the integer of 6-12；Preferably, c val-ala；Preferably, d PABC；PEGn is polyethylene glycol, n indicates poly-
It is right.
Further, formula (13) compound the preparation method comprises the following steps: pectin carboxyl and alcohol condensation after obtain esterification pectin, then with
The C that hydroxyl replaces2-C4Alkylamine generate amide after with two (4- nitrobenzophenone) carbonate reactions up to formula (13) compound.
Further, formula (9) compound the preparation method comprises the following steps: formula (5) compound and adriamycin existing for the alkaline reagent
Under the conditions of react to obtain formula (6) compound, be deprotected under the conditions of formula (6) compound is existing for the piperidines formula (7) compound, again with
R1- b-COOH reacts to obtain formula (8) compound, is deprotected to obtain compound (9) under the conditions of formula (8) compound is existing for the piperidines；Formula
(5) compound is R1- c-PABC, formula (6) compound are R1- c-PABC-e, formula (7) compound are c-PABC-e, formula (8) chemical combination
Object is R1- b-c-PABC-e, R1For blocking group；Preferably, R1For fluorenylmethyloxycarbonyl (Fmoc).
Further, formula (5) compound the preparation method comprises the following steps: formula (3) compound in the presence of acid binding agent with to amino Bian
Alcohol reacts to obtain formula (4) compound, under the conditions of formula (4) compound is existing for the alkaline reagent with two (4- nitrobenzophenone) carbonic esters
Reaction obtains formula (5) compound；Formula (3) compound is R1- c-OH, formula (4) compound are R1- c-PABOH, PABOH are 4- (hydroxyl
Methyl) phenyl amino.
Further, formula (3) compound the preparation method comprises the following steps: formula (1) compound reacts in the presence of NHS and DCC
Formula (2) compound is obtained, formula (2) compound reacts to obtain formula (3) compound with amino acid in the presence of alkaline reagent；Formula (1)
Compound is R1-c1- OH, formula (2) compound are R1-c1- OSu, c1For amino acid；Preferably, c1For val.
The invention also discloses the use that pectin as described above-adriamycin conjugates is used to prepare the drug for the treatment of cancer
On the way.
Advantages of the present invention and effect:
Pectin of the invention-adriamycin conjugates has completely new chemical structure, being capable of the accumulation for a long time of targeting high concentration
In in malignant tumor tissue, achieving the purpose that Synergy and attenuation, indication is the chemotherapy of various solid malignants.
Detailed description of the invention
Fig. 1 is compound 61H NMR spectra；
Fig. 2 is compound 71H NMR spectra；
Fig. 3 is compound 81H NMR spectra；
Fig. 4 is compound 813C NMR spectra；
Fig. 5 is compound 91H NMR spectra；
Fig. 6 is compound 131H NMR spectra；
Fig. 7 is final product adriamycin-pectin conjugates1H NMR spectra；
For final product adriamycin-pectin conjugates, (pectin and adriamycin directly pass through amido bond to Fig. 8 with control compound
Bonding) to the inhibiting effect of H22 liver cancer mouse tumor model；
For final product adriamycin-pectin conjugates, (pectin and adriamycin directly pass through amido bond to Fig. 9 with control compound
Bonding) to the inhibiting effect of 4T1 breast cancer mouse tumor model.
Specific embodiment
By the following examples, the present invention is described in further detail, but these embodiments not to the present invention have appoint
What is limited.
Embodiment 1
150g compound 1 (0.44mol) is dissolved in 1.5LTHF, sequentially add NHS (56g, 0.49mol), DCC (137g,
0.66mol), it is stirred overnight at room temperature, filters, solid methylene chloride is washed twice, and filtrate is concentrated to dryness, and is recrystallized primary
Obtain 2 net product of compound.
Embodiment 2
147g compound 2 (0.34mol) is dissolved in 1.5LTHF, sequentially adds l-Alanine (32g, 0.35mol), NaHCO3
(30g, 0.35mol) adds 500mL water.The ratio for adjusting THF and water makes reaction solution at single-phase (more difficult).It was stirred at room temperature
Night, end of reaction, concentration remove THF, are diluted with water, and HCl adjusts PH to 3-4, and solid, solid washing, vacuum drying, second is precipitated
Acetoacetic ester washing, obtains 130g product 3 (containing a little impurity), crystallization purifying.
Embodiment 3
30g compound 3 (73mmol) is suspended in methylene chloride 500mL, sequentially adds 4- aminobenzyl alcohol (11g, 90mmol),
EEDQ (27g, 113mmol) is eventually adding methanol until solution clarification, is stirred overnight at room temperature, filters, and solid methylene chloride is washed,
Filtrate is concentrated to dryness to obtain crude Compound 4, then is washed to obtain sterling 20g with methyl tertiary butyl ether(MTBE).
Embodiment 4
21g compound 4 (41mmol) is suspended in 500mL methylene chloride, is sequentially added DIPEA (16g, 126mmol), two
(p-nitrophenyl) carbonic ester (18.5g, 69mmol) is eventually adding DMF until solution clarification, is stirred overnight at room temperature.Vacuum distillation
DMF is removed, thick solid is first washed with a small amount of methanol, then recrystallizes number with petroleum ether and ethyl acetate (or methyl tertiary butyl ether(MTBE))
It is secondary, obtain 7g pure compound 5.
Embodiment 5
It weighs 690mg doxorubicin hydrochloride, 1.036g compound 5 in a round bottom flask, finishes, 10mLDMF is added.Stirring
Lower addition DIPEA 0.21mL reacts at room temperature 5h.Reaction solution is slowly dropped into methyl tertiary butyl ether(MTBE) by end of reaction, and centrifugation obtains
Crude product.It is dissolved again with a small amount of DMF, carries out second and recrystallize.Obtain product 1.076g, yield=78%.
Embodiment 6
It weighs 400mg compound 6 in a round bottom flask, finishes, 10mLDMF is added.It is stirred at room temperature down and rapidly joins piperidines
1.8mL reacts at room temperature 70s.End of reaction quickly pours into reaction solution in the methyl tertiary butyl ether(MTBE) in ice bath, and centrifugation obtains slightly
Product.It is dissolved again with a small amount of DMF, is slowly dropped into methyl tertiary butyl ether(MTBE) at room temperature and carries out second of recrystallization.Obtain product
210mg, yield=66%.
Embodiment 7
It weighs 565mgPEG8 in a round bottom flask, 5mLDMF is added.Finish, be stirred at room temperature lower addition HATU 440mg,
DIPEA 0.13mL.Compound 7 is dissolved with a small amount of DMF, dropper is added in reaction flask and (shares solvent 5mL).It is stirred at room temperature anti-
Answer 1.5h.Reaction solution is slowly added in methyl tertiary butyl ether(MTBE) by end of reaction, and centrifugation obtains crude product.It is dissolved again with a small amount of DMF, then
It carries out recrystallizing for second in secondary instillation methyl tertiary butyl ether(MTBE).Column chromatographic purifying: Ea:MeOH=20:1.Yield 55%.
The confirmation of compound 8:
1H NMR(400MHz,DMSO-d6)ppm:14.03(s,1H),13.27(s,1H),9.88(s,1H),8.16(d,j
=6.8Hz, 1H), 7.91-7.87 (m, 5H), 7,85-7.63 (m, 3H), 7,54 (d, j=8.4Hz, 2H), 7.51 (t, j=
7.6Hz, 3H), 7.32 (t, j=7.6Hz, 2H), 7.24 (d, j=8.4Hz, 2H), 6.19 (s, 1H), 5.45 (s, 1H), 5.22
(d, j=2.4Hz, 1h), 4.9 (m, 4H), 4.57 (s, 2H), 4.38-4.15 (m, 8H), 4.0 (s, 3H), 3.6 (t, j=
6.8Hz,2H),3.5-3.3(m,32H),3.12(m,2H),2.96(m,2H),2.42(m,2H),2.2(m,2H),1.9(m,
2H), 1.43 (m, 1H), 1.22 (m, 4H), 0.86 (d, j=4Hz, 3H), 0.82 (dj=4Hz, 3H)
13C NMR(100MHz,DMSO-d6)ppm:171.11,170.91,170.40,160.78,156.21,156.13,
155.36,154.54,143.93,140.76,128.56,127.64,127.09,125.19,120.13,118.92,110.60,
74.98,69.80,69.72,69.61,69.51,69.12,66.94,65.37,63.76,57.49,56.58,49.05,
46.75,35.91,10.61,19.18,18.14,17.93,17.07.
MALDI-TOPMS:m/z 1531.2103[(M+Na+H)+,C77H97N5O26].
RP-HPLC (Agilent 1260) 20.641min, peak area: 92.4038%.
Embodiment 8
It weighs 200mg compound 8 in a round bottom flask, finishes, 6mLDMF is added.It is stirred at room temperature down and rapidly joins piperidines
660uL reacts at room temperature 140s.End of reaction quickly pours into reaction solution in the methyl tertiary butyl ether(MTBE) in ice bath, and centrifugation obtains slightly
Product.It is dissolved again with a small amount of DMF, is slowly dropped into methyl tertiary butyl ether(MTBE) at room temperature and carries out second of recrystallization.
Embodiment 9
Take citrus pectin (raw material of purchase) 2g to be dissolved in 200mL (0.2mol/L) dilute nitric acid solution, be warming up to 85 degree it is (interior
83 degree of temperature, 90 degree of outer temperature), it is warming up to 85 degree (interior temperature is 83 degree practical) and starts timing, stop heating after reacting about 2h, be cooled to room
Reaction solution is added dropwise in the dehydrated alcohol of 800mL by Wen Hou, and film filtering, washing is dried in vacuo to obtain compound 10.
The confirmation of compound 10:
IR(KBr,v,cm-1): 3421 (O-H), 2925 (C-H), 1740 (COOH), 1632 (COOH), 1414,1384,
1334,1234,1146 (C-O-C), 1102 (C-O-C, C-OH), 1018 (C-OH), 952,885,831 (α-D-GalA), 742,
634.
1H NMR(400MHz,D2O)δppm:5.10(s,1H,H-1),5.07(s,1H,H-5))4.50(s,1H,H-4),
4.05 (d, J=10.4,1H, H-3), 3.78 (d, J=10.4, H-2)
13C NMR(100MHz,D2O) δ ppm:172.56 (COOH), 99.86 (C-1, α types), 78.30 (C-5), 70.34
(C-4),68.10(C-3),67.91(C-2).
1H NMR (400MHz, DMSO-d6) δ ppm:12.52 (COOH, 1H), 5.04-4.85 (m, 3H), 4.16 (s, 1H),
(3.67-3.59 m, 3H)
13C NMR (100MHz, DMSO-d6) δ ppm:170.44 (COOH), 98.69 (C-1, α types), 76.71 (C-5),
69.55 (C-4), 68.24 (C-3), 67.90 (C-2)
Embodiment 10
It takes 2g compound 10 in single port bottle, the methanol of 20mL and the dense HCl of 2mL is added, interior temperature is warming up to 55 degree and starts
Timing stops stirring after reacting 7h, and reaction solution is added dropwise in dehydrated alcohol by cooling, is stirred, and is filtered, and washing is dried in vacuo
Compound 11.
Embodiment 11
The compound 11 of 600mg is added into round-bottomed flask, adds 3- amino -1- propyl alcohol (the thick liquid of 3.16g
Body), it stirs 2 days, the reaction time is more than 48h, and clear thick reaction solution is slowly added dropwise in dehydrated alcohol, is stirred, mistake
Filter, washing, is dried in vacuo to obtain compound 12.
Embodiment 12
It takes the compound 12 of 1g to be dissolved in the DMSO of 15mL to make it completely dissolved, adds the two (p-nitrophenyls of 1.29g
Base) carbonic ester, the DIEA of 0.7mL is added, reaction overnight (reaction is more than 12h) finally reaction solution is slowly added dropwise molten into EA
It in liquid (200mL), quickly filters, washing is dried in vacuo to obtain compound 13.
The confirmation of compound 13:
1H NMR (400MHz, DMSO-d6) δ ppm:8.305 (s, 2H), 7.56 (d, 2H), 4.88-4.79 (m), 4.26-
4.02 (m), 3.64-3.20 (m), 1.98 (s, 2H), 1.58 (s, 1H), 1.19-1.16 (m)
Embodiment 13
Weigh Compound 13 is completely dissolved in the mixed solution of DMF and DMSO, compound 9 is added, after mixing evenly
DIEA is added, room temperature reaction for 24 hours, then directly uses solution DMSO- water gradient to dialyse, and HPLC is detected to no adriamycin and its spread out
Pectin-adriamycin conjugates is lyophilized to obtain in biology after the completion of dialysis.
The confirmation of pectin-adriamycin conjugates:
1H NMR (400MHz, DMSO-d6) δ ppm:14.03 (s, 1H), 13.26 (s, 1H), 9.89 (s, 1H), 8.31-
6.82 (m, 10H), 5.47-3.38 (m), 1.95-0.84 (m)
Embodiment 14
Pectin-adriamycin conjugates (can be prepared by embodiment 1-13) has the following structure in the present embodiment:
Vitro cytotoxicity test: collect logarithmic phase growth HT-29 colon cancer cell, HepG-2 liver cancer cells,
SMMC7721 liver cancer cells, SKOV3 ovarian cancer cell and MCF-7 breast cancer cell, adjust concentration of cell suspension, and every hole is added
100 μ L, bed board make 4000/hole of cell density to be measured (edge hole is filled with sterile PBS).5%CO2, 37 DEG C are incubated for for 24 hours,
Culture medium is discarded, concentration gradient (being 0.125,0.25,0.5,1.0,2.0,4.0,8.0 and 16.0 μ g/mL containing adriamycin) is added
Pectin adriamycin or adriamycin, each concentration sets 3 multiple holes.Then in 5%CO2, 48h is incubated in 37 DEG C of incubator.Every hole
Continue to cultivate 2h after 10 μ l CCK-8 solution are added.With the absorbance in microplate reader measurement each hole at 450nm, pectin Ah mould is calculated
The IC of element and adriamycin to various cancer cells50Value.
1 vitro cytotoxicity test of table
Vitro cytotoxicity test shown in table 1 the result shows that, the pectin that the present invention is prepared-adriamycin conjugates pair
The inhibiting rate and positive control drug adriamycin of various cancer cells have preferable Vitro Cytotoxicity without significant difference.
Embodiment 15
In vivo due to the pectin-adriamycin conjugates (can be prepared by embodiment 1-13) of the invention being prepared
It is by the accumulation of EPR effect in tumor tissues, achieving the purpose that passive target, therefore investigates the fruit that the present invention is prepared
Internal pharmacodynamic results of the glue-adriamycin conjugates to tumor-bearing mice.
Internal pharmacodynamic test: H22 liver cancer cells, the 4T1 breast cancer cell, EMT6 breast cancer cell of logarithmic growth phase are collected
And HT-29 colon cancer cell, adjustment concentration of cell suspension are 3 × 107A cell/mL, by cell inoculation in Balb/c mouse or
The subcutaneous 0.1mL/ of nude mice right upper extremity is only (containing about cell number 3 × 106It is a), good mouse tumor volume average out to 100mm to be seeded3When
Mice with tumor is grouped at random, respectively negative control group (0.9% sodium chloride injection), adriamycin control group (5mg/kg), fruit
Glue adriamycin experimental group (adriamycin equivalent is 5mg/kg), every group of 8 mouse are injected intravenously above-mentioned various drugs, 4 are administered altogether
It is secondary.Record tumor volume change and weight, volume calculation formula are as follows: volume=(length x width2)/2。
In vivo in effect experiment, pectin-adriamycin conjugates that the present invention is prepared is to H22 liver cancer, 4T1 mammary gland
The curative effect of cancer, EMT-6 breast cancer and HT-29 colon cancer is superior to adriamycin control.Pectin-adriamycin that the present invention is prepared
Conjugates can obviously inhibit H22 liver cancer, 4T1 breast cancer, the tumour growth of EMT-6 breast cancer tumor-bearing mice, and inhibiting rate is higher,
Respectively 72.98%, 78.1%, 71.68%；And the adriamycin as control, to H22 liver cancer, 4T1 breast cancer, EMT-6 mammary gland
The inhibiting rate of cancer tumor-bearing mice is respectively as follows: lower than pectin-adriamycin conjugates inhibiting rate that the present invention is prepared
53.42%, 50.61%, 56.95%.In test, the pectin that the present invention is prepared-adriamycin conjugates group mouse body
Weight nothing is decreased obviously, no dead mouse；And the decline of adriamycin group mouse weight is obvious, weight loss is more than 15%, and is had a
Other mouse occurs dead.Internal pharmacodynamic results show that compared with adriamycin, pectin-adriamycin that the present invention is prepared is conjugated
Object has the function of Synergy and attenuation.Tumor tissues small animal living body fluorescence imaging system is taken after H22 liver cancer tumor-bearing mice is put to death
Overall view examines the adriamycin image intensity in tissue, and pectin-adriamycin conjugates administration group that the discovery present invention is prepared is small
Mouse tumour shows adriamycin hyperfluorescence, illustrates that pectin-adriamycin conjugates that the present invention is prepared can long-time, high concentration
Accumulation in tumor tissues.
Embodiment 16
In order to the pectin being prepared-adriamycin conjugates more of the invention (can be prepared by embodiment 1-13) with
The curative effect of existing control compound (pectin directly passes through amido bond with adriamycin and is bonded), we select H22 liver cancer and 4T1 cream
Gland cancer mouse model is compared.
(pectin and adriamycin pass through amide for the pectin that 2 present invention of table is prepared-adriamycin conjugates and control compound
Key bonding) nature difference compare
The result shows that pectin-adriamycin conjugates that the present invention is prepared is swollen to H22 liver cancer and 4T1 breast cancer mouse
The inhibiting rate of tumor model is above existing control compound (pectin directly passes through amido bond with adriamycin and is bonded), inhibiting rate point
Not Wei 70.83%, 76.20%, and the inhibition of existing control compound (pectin directly passes through amido bond with adriamycin and is bonded)
Rate only has 48.64%, 47.00%.As can be seen from figures 8 and 9, inventor also found in an experiment, the fruit that the present invention is prepared
Glue-adriamycin conjugates is not only high to the inhibiting rate of tumour, can also maintain higher inhibiting rate for a long time, that is, give identical
Dosage, the inhibiting effect time that pectin-adriamycin conjugates maintains are longer.
Simply to illustrate that technical concepts and features of the invention, its purpose is allows in the art above-described embodiment
Those of ordinary skill cans understand the content of the present invention and implement it accordingly, and the present invention and synthetic route optimization can not be limited with this
Protection scope.Equivalent changes or modifications, should all cover in the present invention made by the essence of all contents according to the present invention
Protection scope in.
Claims (10)
1. a kind of pectin-adriamycin conjugates, which is characterized in that have such as formula (I) structure:
a-b-c-d-e(I)
Wherein a is pectin or modifying pectin, and b is polyethylene glycol PEGn, and c is enzymatic lysis group, polypeptide or polypeptide derivative, d
For self-degradation group, e is adriamycin.
2. pectin as described in claim 1-adriamycin conjugates, which is characterized in that the pectin is polygalacturonase
Content is 90% or more, molecular weight is 1-60KD without ester pectin, and the modifying pectin is polygalacturonase without ester pectin
Carbonyl and hydroxyl replace C2-C4Alkylamine generate amide, be further activated as pectin carbonic ester formula (13)；It is preferred that
, a is modifying pectin；Further preferably, the molecular weight of a is 1-4KD；Further preferably, the polygalacturonase is without ester
The galacturonic acid content of pectin is 95% or more.
3. pectin as described in claim 1-adriamycin conjugates, which is characterized in that in formula (I), n is the integer of 1-20, and c is
Any one in dipeptides, dipeptidase derivant, tripeptides, tripeptide derivative, tetrapeptide or tetrapeptide derivative；Preferably, n is 6-12's
Integer；Preferably, c val-ala.
4. pectin as described in claim 1-adriamycin conjugates, which is characterized in that d is self-degradation group amino benzyloxy carbonyl
Base；Preferably, the aminobenzyloxycarbonyl is to aminobenzyloxycarbonyl, i.e. PABC.
5. a kind of preparation method of pectin according to any one of claims 1-4-adriamycin conjugates, which is characterized in that will
It is reacted under the conditions of formula (13) compound and formula (9) compound are existing for the alkaline reagent,
In formula (13), a is the integer selected from 2-4, m 1-60KD；Formula (9) compound is b-c-d-ADM, b PEGn；C is more
Peptide, it is preferred that c is any one in dipeptides, dipeptidase derivant, tripeptides, tripeptide derivative, tetrapeptide or tetrapeptide derivative；D is
Self-degradation group, it is preferred that n is the integer of 6-12；Preferably, c val-ala；Preferably, d PABC.
6. pectin as claimed in claim 5-adriamycin conjugates preparation method, which is characterized in that formula (13) compound
The preparation method comprises the following steps: pectin obtains esterification pectin, then the C replaced with amino after reacting with alcohol2-C4Alcohol reaction generate amide after again
With two (4- nitrobenzophenone) carbonate reactions up to formula (13) compound.
7. pectin as claimed in claim 5-adriamycin conjugates preparation method, which is characterized in that the system of formula (9) compound
Preparation Method are as follows: react to obtain formula (6) compound, formula (6) chemical combination under the conditions of formula (5) compound and adriamycin are existing for the alkaline reagent
Be deprotected under the conditions of object is existing for the piperidines formula (7) compound, (7) compound again with R1- b-COOH reacts to obtain formula (8) chemical combination
Object is deprotected to obtain compound (9) under the conditions of formula (8) compound is existing for the piperidines；Formula (5) compound is R1- c-PABC, formula
(6) compound is R1- c-PABC-e, formula (7) compound are c-PABC-e, and formula (8) compound is R1- b-c-PABC-e, R1To protect
Protect group；Preferably, R1For fluorenylmethyloxycarbonyl Fmoc.
8. pectin as claimed in claim 7-adriamycin conjugates preparation method, which is characterized in that the system of formula (5) compound
Preparation Method are as follows: formula (3) compound in the presence of acid binding agent with aminobenzyl alcohol alcohol is reacted to obtain formula (4) compound, formula (4) chemical combination
Formula (5) compound is obtained with two (4- nitrobenzophenone) carbonate reactions under the conditions of object is existing for the alkaline reagent；Formula (3) compound
For R1- c-OH, formula (4) compound are R1- c-PABOH, wherein PABOH is 4- (methylol) phenyl amino；C is polypeptide, preferably
, c val-ala.
9. pectin as claimed in claim 8-adriamycin conjugates preparation method, which is characterized in that formula (3) compound
The preparation method comprises the following steps: formula (1) compound reacts in the presence of NHS and DCC obtains formula (2) compound, formula (2) compound is in alkalinity
It reacts to obtain formula (3) compound with amino acid in the presence of reagent；Formula (1) compound is R1-c1- OH, formula (2) compound are R1-
c1- OSu, c1For amino acid；Preferably, c1For val.
10. the use that a kind of pectin according to any one of claims 1-4-adriamycin conjugates is used to prepare treating cancer drug
On the way.
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